Urine analysis
urine
analysis
Normal Pathological
constituents constituents
inorganic
organic
Composition of urine
Its depend upon kidney functions
− Glomerular filtration
− Tubular reabsorption
− Tubular secretion
Normally urine contains 90-95 % (1500ml/day) water and 5-10% solid material (60gm).
They may be organic and inorganic
Organic constituents of urine
− Urea 20-30gm
− Urine acid 0.5-0.8gm
− Hippuric acid 0.7-0.8 gm
− Creatinine 1-1.8 gm
− Amino acid 0.5-1gm
Inorganic constituents
− Cl‾ 10-15 gm
− PO4‾³ 1-1.5gm
− SO4‾² 1-1.2gm
− Ca⁺² 0.1-0.3 gm
Examination of urine
For detail read chapter urine analysis from book
− Physical
− Chemical
− Microscopic
Physical examination
Colour amber(urochrome)
Volume 800-2600ml (1500ml/day)
pH 4.58-6.0
Odour aromatic or ammonical
Specific gravity 1.003-1.030
Object
To analyse the normal inorganic constituents of urine
Inorgani constituents of urine
Experiment Observation Inference
1. Test for chloride White ppt. of AgCl Cl⁻ is present
Take 3 ml of urine sample add 3 obtained
drops of concentrated HNO3
and 1 ml of AgNO3.
2. Test for sulphate White ppt. of BaSO4 Sulphate is present
Take 3 ml of urine sample add 3 formed
drops if concentrated HCl and
few drops of BaCl2
3. Test for phosphate Yellow ppt. obtained Phosphate is present
Take 3 ml of urine sample add
1ml of nitric acid and 2 ml of
ammonium molybdate and heat
it.
4. Test for calcium White ppt of Calcium Calcium is present
Take 5 ml of urine sample add 2 oxalate is formed
ml of sulkowitch reagent drop
wise
Importance;
• Urine normally contains 6.0 to 16.0 gm of NaCl per litre.High levels of
urine chloride may indicate dehydration, fasting, adrenal gland failure
(Addison's disease) or increased dietary salt intake.
• Urinary sulfate is a reflection of dietary protein intake, particularly of
meat.
• High phosphate/phosphorous levels in urine may be due to Kidney
disease and Hyperparathyroidism.
• Urine calcium levels are too high or too low, it may be a sign of kidney
disease, kidney stones, bone disease, a parathyroid gland disorder, or
other conditions.
Object
To analyse the normal inorganic constituents of urine
Experiment Observation Inference
1. Test for urea Violet or purple urea is present
colour appear
(Biuret test)
Take few crystal of urea add 3mlof 5%
NaOH and add I ml of CuSO4 and 1 ml
of original solution and heat it.
2. Tests for creatinine I. Red colour Creatinine is
I. Jaffe’s Test(picric acid test) formed present
Take 5 ml of urine sample add 5ml of II. Red colour
aqueous solution of picric acid and add turns yellow on
1ml of 10%NaOH. addition of
II. Nitroprusside test NaOH
III. A green colour
Take 5 ml of urine sample and add few formed finally
drops of sodium Nitroprusside and few it will turns
drops of 10% NaOH into blue
III. Nitroprusside acetic acid test colour solution
To the yellow solution or ppt obtained or ppt.
from the above test add an excess of
acetic acid and heat the solution
3. Test for uric acid I. Reddish Uric acid is
i. Murexide test residue first present
Take few crystals of uricacid add 2-3 obtained has
drops of concentrated HNO3 crefully turn to
(evaporate to dryness or heat on a purplish red
water bath till a reddish residue occur, solution .the
cool it under tap water then add one colour is due
drop of dilute NH4OH solution to the
formation of
ii. Fehling’s test murexide
Take few crystal of uric acid add 3-4 ml
of Na2CO3 solution boil and add 2.5 ml
of Fehling’s solution A and B then boil
it .
II. red ppt.
obtained
Uric acid is
present
4. Test for hippuric acid Green or orange ppt Hippuric acid is
Take 5 ml of urine and few drops of obtained present
KOH and few drops of ferric chloride.
Importance of normal constituents of urine
1. Biuret test
Urea is a nitrogenous end product of the metabolic breakdown of proteins. Biuret is a
compound formed by heating urea to 180'C. When biuret is treated with dilute copper
sulfate in alkaline medium, a purple colour is obtained. Biuret test is answered by
compounds containing two or more CO-NH groups.
2. Jaffe’s Test(picric acid test)
Creatinine is a break-down product of creatine phosphate in muscle tissue. Creatinine
reacts with alkaline picrate to form Creatinine picrate, an orange red coloured complex.
3. Test for uric acid
Murexide test
Uric acid is oxidized by nitric acid to give purpuric acid (reddish yellow).This in
turn combines with ammonia to form purple red. Uric acid is a normal body waste product. It
forms when purines break down.
4. Test for hippuric acid
Hippuric acid is normally present in human urine as a metabolite of dietary components.
Therefore, the analysis of hippuric acid in urine provides an diet and liver diagnostic test .
In humans the synthesis of hippuric acid takes place almost entirely in the liver when
benzoic acid combines with glycine.
Object
To analyse the abnormal/pathological constituents of urine
Experiment Observation Inference
1. Test for protein White ppt formed protein is present
I. Sulphosalicylic test
Take 2 ml of urine sample in a
test tube add few drops of 20%
Sulphosalicylic acid and shake.
II. Coagulation Test Coagulum formed Protein is present
Heat 5ml of urine sample
2. Test for sugars Red ppt. formed Reducing sugar is
I. Benedict’s test present
Take 5 ml of qualitative
Benedict’s reagent 1ml of urine
sample and boil
3. Test for ketone bodies Permanganate colour Ketone bodies are
Rothera’s test appear present
5ml of urine sample add excess
of ammonium sulphate shake it
.add 4-5 drops of 2% sodium
Nitroprusside and add 2 ml of
concentrated ammonia
solution. Allow to stand.
4. Test for bile pigments Green blue colour appear Bile pigments are
Fouchet’s test present
5ml of urine sample add few
drops of acetic acid, saturated
ammonium sulphate and 2ml
of 10% barium chloride. Mix
well and flter, to the ppt. add 2
drops of fouchet’s reagent.
5. Test for bile salts Sulphur particles sinks Bile salt is present
Hay’s test down
Take 2ml of urine sample in
attest tube and sprinkle a little
finely powdered sulphur on the
surface.
6. Test for blood Green or blue colour Blood is present
Benzidine test appear
1ml saturated solution of
benzidine in a glacial acetic
acid add 3ml add 1ml
hydrogen peroxide.
Principles of tests
Test for ketone bodies
Rothera’s test
Nitroprussidien alkalinem edium reacts with keto group of ketone bodies (acetone
and acetoacetateto) form a purple ring.
Test for bile pigments
Fouchet’s test
Bile pigments are adsorbed on barium sulfate. Fouchet's reagents (containing ferric
chloride in trichloroacetic acid) oxidizes bilirubin to biliverdin (green) and bilicyanin
(blue).
Test for blood
Benzidine test
Hemoglobin (acts like peroxidase) decomposes hydrogen peroxide to liberate nascent
oxygen (O-) which oxidises benzidine to a green or blue coloured complex.
